Retz M, Sidhu S S, Lehmann J, Tamamura H, Fujii N, Basbaum C
Biomolecular Sciences Program, Cardiovascular Research Institute, Department of Anatomy, UCSF, San Francisco, CA 94143-0452, USA.
Eur Urol. 2005 Dec;48(6):1025-30. doi: 10.1016/j.eururo.2005.07.016. Epub 2005 Aug 15.
The CXCR4/CXCL12 axis appears crucial in the metastasis of bladder cancer. Our aim was to evaluate the potency of the CXCR4 antagonist, 4F-benzoyl-TE14011 (4F-bTE), as an anti-metastatic drug in this disease. In this study, we assessed the ability of 4F-bTE to inhibit tumor cell motility, invasion through extracellular matrix (ECM), matrix metalloproteinase (MMP) secretion and cytoskeletal responses to chemokine.
To assess the degree to which cells could migrate and invade ECM under various conditions, we used TCCSUP bladder cancer cells in a Boyden chamber system. To monitor actin polymerization, we stained cells on chamber slides with AlexaFluor 594 phalloidin. To measure matrix-metalloproteinase-2 and -9 (MMP) activity, we used gelatin zymography. To assess the effects of the CXCR4 antagonist 4F-bTE on each of the above parameters, we exposed bladder cancer cells either to chemokine CXCL12, alone, or to both CXCL12 and 4F-bTE. We also monitored cells for apoptotic and necrotic changes during drug treatment.
The CXCR4 antagonist 4F-bTE markedly decreased CXCL12-induced bladder cancer cell migration and ECM invasion in Boyden chamber assays. The antagonist also blocked chemokine-induced actin polymerization as well as the induction of MMP-2 and MMP-9 in these cells.
The CXCR4 antagonist 4F-bTE has the potential to inhibit expression of the metastatic phenotype and may provide therapeutic value to patients.
CXCR4/CXCL12轴在膀胱癌转移中似乎至关重要。我们的目的是评估CXCR4拮抗剂4F-苯甲酰基-TE14011（4F-bTE）作为该疾病抗转移药物的效力。在本研究中，我们评估了4F-bTE抑制肿瘤细胞运动、通过细胞外基质（ECM）侵袭、基质金属蛋白酶（MMP）分泌以及细胞骨架对趋化因子反应的能力。
为了评估细胞在各种条件下迁移和侵袭ECM的程度，我们在Boyden小室系统中使用了TCCSUP膀胱癌细胞。为了监测肌动蛋白聚合，我们用AlexaFluor 594鬼笔环肽对小室载玻片上的细胞进行染色。为了测量基质金属蛋白酶-2和-9（MMP）的活性，我们使用了明胶酶谱法。为了评估CXCR4拮抗剂4F-bTE对上述每个参数的影响，我们将膀胱癌细胞单独暴露于趋化因子CXCL12，或同时暴露于CXCL12和4F-bTE。我们还监测了药物治疗期间细胞的凋亡和坏死变化。
在Boyden小室试验中，CXCR4拮抗剂4F-bTE显著降低了CXCL12诱导的膀胱癌细胞迁移和ECM侵袭。该拮抗剂还阻断了趋化因子诱导的肌动蛋白聚合以及这些细胞中MMP-2和MMP-9的诱导。
CXCR4拮抗剂4F-bTE具有抑制转移表型表达的潜力，可能为患者提供治疗价值。
